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Estradiol treatment induces testicular oxidative stress and germ cell apoptosis in rats

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Abstract

In order to understand the pathogenesis of estradiol induced effects in the seminiferous epithelium, studies were undertaken in adult rats with estradiol-3-benzoate administered for different durations. After 30 d of treatment, a significant rise in lipid peroxidation with concomitant fall in the activities of superoxide dismutase and catalase was observed. Both, serum and intra-testicular testosterone levels were found severely depleted. Seminiferous epithelium was devoid of elongated spermatids and spermatozoa by 30 d of treatment. Number of spermatocytes and round spermatids were significantly (p < 0.001) reduced. Flowcytometric analysis confirmed a drastic reduction of the haploid cell population (1c peak). Beginning from day 10 of treatment, there was a consistent rise in the number of pyknotic/apoptotic germ cells in the seminiferous epithelium. A gradual increase in Bax protein expression was observed with the duration of treatment. The shift in Bax immunostaining from the cytoplasm and nucleus of germ cells (at 10 d of treatment) to only nuclei of cells by 30 d of treatment was also noticed. By this time testicular tissue showed three-fold increase in caspase-8 enzyme activity. Viable testicular cells isolated in vitro decreased drastically subsequent to different periods of estradiol treatment. The above findings substantiate the fact that the testicular pathogenesis of estradiol benzoate treatment may be primarily because of altered reproductive hormone levels and high oxidative stress leading to germ cell apoptosis and subsequent germ cell loss in the seminiferous epithelium.

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References

  1. Sharpe RM (1998) Roles of estrogen in the male. Trends Endocrinol Metab 9:371–377

    Article  CAS  PubMed  Google Scholar 

  2. Lubahn DB, Moyer JS, Golding TS, Course JF, Korach KS, Smithies O (1993) Alteration of reproductive function but not prenatal sexual development after insertional disruption of the mouse estrogen receptor gene. Proc Natl Acad Sci USA 90:11162–11166

    Article  PubMed  CAS  Google Scholar 

  3. Korach KS (1994) Insights from the study of animals lacking functional estrogen receptor. Science 266:1524–1527

    PubMed  CAS  Google Scholar 

  4. Eddy EM, Washburn TF, Bunch DO, Goulding EH, Gladen BC, Lubahn DB, Korach KS (1996) Targeted disruption of the estrogen receptor gene in male mice causes alteration of spermatogenesis and infertility. Endocrinology 137:4796–4805

    Article  PubMed  CAS  Google Scholar 

  5. Robertson KM, O'Donnell L, Jones ME, Meachem SJ, Boon WC, Fisher CR, Graves KH, McLachlan RI, Simpson ER (1999) Impairment of spermatogenesis in mice lacking a functional aromatase (Cyp 19) gene. Proc Natl Acad Sci USA 96:7986–7991

    Article  PubMed  CAS  Google Scholar 

  6. Hess RA, Bunick D, Lee KH, Bahr J, Taylor JA, Korach KS, Lubahn DB (1997) A role for estrogen in the male reproductive system. Nature 390:509–512

    Article  PubMed  CAS  Google Scholar 

  7. Lee KH, Hess RA, Bahr J, Lubahn DB, Taylor J, Bunick D (2000) Estrogen receptor α has a functional role in the mouse rete testis and efferent ductules. Biol Reprod 63:1873–1880

    Article  PubMed  CAS  Google Scholar 

  8. Ebling FJ, Brooks AN, Cronin AS, Ford H, Kerr JB (2000) Estrogenic induction of spermatogenesis in the hypogonadal mouse. Endocrinology 141:2861–2869

    Article  PubMed  CAS  Google Scholar 

  9. Luconi M, Muratori M, Forti G, Baldi E (1999) Identification and characterization of a novel functional estrogen receptor on human sperm membrane that interferes with progesterone effects. J Clin Endocrinol Metab 84:1670–1678

    Article  PubMed  CAS  Google Scholar 

  10. Pentikainen V, Erkkila K, Suomalainen L, Parvinen M, Dunkel L (2000) Estradiol acts as a germ cell survival factor in the human testis in vitro. J Clin Endocrinol Metab 85:2057–2067

    Article  PubMed  CAS  Google Scholar 

  11. Pelletier G, Labrie C, Labrie F (2000) Localization of oestrogen receptor α, oestrogen receptor β and androgen receptors in the rat reproductive organs. J Endocrinol 165:359–370

    Article  PubMed  CAS  Google Scholar 

  12. Blanco-Rodriguez J, Martinez-Garcia C (1997) Apoptosis pattern elicited by estradiol treatment of the seminiferous epithelium of the adult rat. J Reprod Fertil 110:61–70

    Article  PubMed  CAS  Google Scholar 

  13. Tena-Sempere M, Navarro J, Pinilla L, Gonzalez LC, Huhtaniemi I, Aguilar E (2000) Neonatal exposure to estrogen differently alters estrogen receptor α and β mRNA expression in rat testis during postnatal development. J Endocrinol 165:345–357

    Article  PubMed  CAS  Google Scholar 

  14. De Jong FH, Uilenbroek TJ, Van der Molen HJ (1975) Oestradiol 17β, testosterone and gonadotrophins in oestradiol-17β-treated intact adult male rats. J Endocrinol 65:281–282

    Article  Google Scholar 

  15. Blanco-Rodriguez J, Martinez-Garcia C (1998) Apoptosis precedes detachment of germ cells from the seminiferous epithelium after hormone suppression by short-term estradiol treatment of rats. Int J Androl 21:109–115

    Article  PubMed  CAS  Google Scholar 

  16. O'Donnell L, Robertson KM, Jones ME, Simpson ER (2001) Estrogen and spermatogenesis. Endocr Rev 22:289–318

    Article  PubMed  Google Scholar 

  17. Russell LD, Clermont Y (1977) Degeneration of germ cells in normal, hypophysectomized and hormone treated hypophysectomized rats. Anat Rec 187:347–366

    Article  PubMed  CAS  Google Scholar 

  18. Abercrombie M (1946) Estimation of nuclear population from microtome sections. Anat Rec 94:239–247

    Article  Google Scholar 

  19. Clermont Y, Morgentaler H (1955) Quantitative study of spermatogenesis in the hypophysectomized rat. Endocrinology 57:369–382

    PubMed  CAS  Google Scholar 

  20. Nagao Y (1989) Viability of meiotic prophase spermatocytes of rats is facilitated in primary culture of dispersed testicular cells on collagen gel by supplementing epinephrine or norepinephrine: evidence that meiotic prophase spermatocytes complete meiotic divisions in vitro. In Vitro Cell Dev Biol 25:1088–1098

    PubMed  CAS  Google Scholar 

  21. Spano M, Evenson DP (1993) Flow cytometric analysis for reproductive biology. Biol Cell 78:53–62

    Article  PubMed  CAS  Google Scholar 

  22. Gautam DK, Misro MM, Chaki SP, Sehgal N. H2O2 at physiological concentrations modulates Leydig cell function inducing oxidative stress and apoptosis. Apoptosis 11:39–46

  23. Lowry OH, Rosebrough NJ, Farr AH, Randall RJ (1951) Protein measurement with the folin phenol reagent. J Biol Chem 193:265–275

    PubMed  CAS  Google Scholar 

  24. Ohkawa H, Ohishi N, Yagi K (1979) Assay for lipid peroxides in animal tissues by thiobarbituric acid reaction. Ann Biochem 95:351–358

    Article  CAS  Google Scholar 

  25. Das K, Samantha L, Chainy GBN (2000) A modified spectrophotometric assay of superoxide dismutase using nitrite formation of superoxide radicals. Indian J Biochem Biophys 37:201–204

    CAS  Google Scholar 

  26. Aebi H (1984) Catalase in vitro. Meth Enzymol 105:121–126

    Article  PubMed  CAS  Google Scholar 

  27. Abraham GE (1974) Radioimmunoassay of steroids in biological fluids. Clin Biochem 7:193–201

    Article  PubMed  CAS  Google Scholar 

  28. Cerutti PA (1985) Prooxidant states and tumor promotion. Science 227:375–381

    PubMed  CAS  Google Scholar 

  29. Kappus H (1985) Lipid peroxidation: mechanism, analysis, enzymology and biological relevance. In: Sies H (ed) Oxidative stress. Academic Press, London, pp 273–269

    Google Scholar 

  30. Russel LD, Clermont Y (1977) Degeneration of germ cells in normal, hypophysectomized and hormone treated hypophysectomized rats. Anat Rec 187:347–366

    Article  Google Scholar 

  31. Finkelstein JS, O'Dea LS, Whitcomb RW, Crowley Jr WF (1991) Sex steroid control of gonadotropin secretion in the human male. II. Effects of estradiol administration in normal and gonadotropin-releasing hormone-deficient men. J Clin Endocrinol Metab 73:621–628

    Article  PubMed  CAS  Google Scholar 

  32. Hayes FJ, DeCruz S, Seminara SB, Boepple PA, Crowley Jr WF (2001) Differential regulation of gonadotropin secretion by testosterone in the human male: absence of a negative feedback effect of testosterone on follicle-stimulating hormone secretion. J Clin Endocrinol Metab 86:53–58

    Article  PubMed  CAS  Google Scholar 

  33. Bagatell CJ, Dahl KD, Bremner WJ (1994) The direct pituitary effect of testosterone to inhibit gonadotropin secretion in men is partially mediated by aromatization to estradiol. J Androl 15:15–21

    PubMed  CAS  Google Scholar 

  34. Handelsman DJ, Wishart S, Conway AJ (2000) Oestradiol enhances testosterone-induced suppression of human spermatogenesis. Hum Reprod 15:672–679

    Article  PubMed  CAS  Google Scholar 

  35. Russel LD, Malone JP, Karpas SL (1981) Morphological pattern elicited by agents affecting spermatogenesis by disruption of its hormonal stimulation. Tissue Cell 13:369–380

    Article  Google Scholar 

  36. McLachlan RL, Wreford NG, Meachem SJ, De Kretser DM, Robertson M (1994) Effect of testosterone on spermatogenic cell population in the adult rat. Biol Reprod 6:705–709

    Google Scholar 

  37. Chaki SP, Ghosh D, Misro MM (2003) Simultaneous increase in germ cell apoptosis and oxidative stress under acute unilateral testicular ischemia in rats. Int J Androl 26:319–328

    Article  PubMed  CAS  Google Scholar 

  38. Chaki SP, Misro MM, Ghosh D, Gautam DK, Srinivas M (2005) Apoptosis and cell removal in the cryptorchid rat testis. Apoptosis 10:395–405

    Article  PubMed  CAS  Google Scholar 

  39. Choudhury S, Chainy GB, Mishro MM (2003) Experimentally induced hypo- and hyper-thyroidism influence on the antioxidant defense system in adult rat testis. Andrologia 35:131–140

    Article  PubMed  CAS  Google Scholar 

  40. Sahoo DK, Roy A, Bhanja S, Chainy GB (2005) Experimental hyperthyroidism-induced oxidative stress and impairment of antioxidant defense system in rat testis. Indian J Exp Biol 43:1058–1067

    PubMed  CAS  Google Scholar 

  41. Chainy GBN, Samantray S, Samanta L (1997) Testosterone-induced changes in testicular antioxidant system. Andrologia 29:342–349

    Google Scholar 

  42. Kii N, Adachi N, Liu K, Arai T (2005) Acute effects of 17beta-estradiol on oxidative stress in ischemic rat striatum. J Neurosurg Anesthesiol 17:27–32

    PubMed  Google Scholar 

  43. Urata Y, Ihara Y, Murata H, Goto S, Koji T, Yodoi J, Inoue S, Kondo T (2006) 17-beta-estradiol protects against oxidative stress-induced cell death through the glutathione/glutaredoxin-dependent redox regulation of Akt in myocardiac H9c2 cells. J Biol Chem (ahead of print)

  44. Tam NN, Ghatak S, Ho SM (2003) Sex hormone-induced alterations in the activities of antioxidant enzymes and lipid peroxidation status in the prostate of Noble rats. Prostate 55:1–8

    Article  PubMed  CAS  Google Scholar 

  45. Tapanainen JS, Tilly JL, Vinko KK, Hseuj AJW (1993) Hormonal control of apoptotic cell death in the testis:gonadotropins and androgens as testicular survival factors. Mol Endocrinol 7:643–650

    Article  PubMed  CAS  Google Scholar 

  46. Nair R, Shah C (2003) Diethylstilebestrol induces rat spermatogenic cell apoptosis in vivo through increased expression of spermatogenic cell Fas/FasL expression. J Biol Chem 278:6470–6481

    Article  PubMed  CAS  Google Scholar 

  47. Ikeda M, Kodama H, Fukuda J, Shimizu Y, Murata M, Kumagai J, Tanaka T (1999) Role of radical oxygen species in rat testicular germ cell apoptosis induced by heat stress. Biol Reprod 61:393–399

    Article  PubMed  CAS  Google Scholar 

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Correspondence to M. M. Misro.

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Chaki, S.P., Misro, M.M., Gautam, D.K. et al. Estradiol treatment induces testicular oxidative stress and germ cell apoptosis in rats. Apoptosis 11, 1427–1437 (2006). https://doi.org/10.1007/s10495-006-8761-4

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